Pharmaceutical compositions comprising an aromatse inhibitors

ABSTRACT

The present invention relates to low-dose pharmaceutical compositions comprising the aromatase inhibitor 4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile, as the active ingredient in a suitable carrier. The present invention also relates to a process for their preparation and to their use as medicaments.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of application Ser. No. 14/342,813,filed Mar. 5, 2014, which is the National Stage of InternationalApplication No. PCT/US202/053846, filed Sep. 6, 2016, which claims thebenefit of U.S. Provisional Application No. 61/532,459, filed Sep. 8,2011, and U.S. Provisional Application No. 62/638,588, filed Apr. 26,2012, each of which is hereby incorporated in its entirety including alltables, figures and claims.

FIELD OF THE INVENTION

The present invention relates to low-dose pharmaceutical compositionscomprising the aromatase inhibitor4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile, as theactive ingredient in a suitable carrier. The present invention alsorelates to a process for their preparation and to their use asmedicaments.

BACKGROUND OF THE INVENTION

The aromatase inhibitor4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile, alsoknown as4-[α-4-Cyanophenyl)-α-fluoro-1-(1,2,4-triazolyl)methyl]-benzonitrile orCGP47645, first described in 1992 [EP 490 816 and U.S. Pat. No.5,637,605], has the following structural formula (I)

The compound4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile is acrystalline compound with a sharp melting endotherm at 169.5° C. Thecrystalline powder is not hygroscopic and is poorly soluble in water.

4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645) is a highly specific and potent aromatase inhibitor which wasshown here within to have a longer half life in humans than doesletrozole (Femara®), a marketed aromatase inhibitor to which CGP47645 isstructurally related. In vitro experiments with human placentalmicrosomal aromatase demonstrated an IC₅₀=6 nM. Oral administration ofCGP47645 to rats demonstrated a T½ of 75 hours. The exposure expressedas AUC was proportional to the administered dose. In two differentaromatase dependent experimental models, inhibition ofandrostenedione-induced uterine hypertrophy in rats and inhibition ofDMBA-induced mammary tumors in rats, the ED₅₀ was 0.003 mg/kg and 0.01mg/kg, respectively. These results suggested CGP47645 is approximately10-fold more potent than letrozole.

4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile is ahighly potent and cohesive drug which needs to be dispensed in lowdoses. Such drugs require a careful formulation and production in orderto produce solid oral dosage forms with acceptable content uniformityand physical stability. There is a need to formulate4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile intopharmaceutical compositions, especially solid oral dosage forms, suchthat the therapeutic benefits of the compound may be delivered safely toa patient in need thereof.

Formulation of low dose medicines can be very challenging and problemsrelated to content uniformity and physical stability may arise. Contentuniformity is a key parameter for oral solid dosage forms, becausesignificant deviations in active content may impact the performance ofthe product in terms of efficacy and safety. The selection of theexcipients and the specific steps during the manufacturing are criticalfactors and need to be controlled in order to get a homogenous andsegregation-free low dose formulation.

Accordingly, the present invention provides a solid pharmaceuticalcomposition suitable for oral administration, comprising CGP47645.

SUMMARY OF THE INVENTION

In one aspect, the present invention is directed to a low-dosepharmaceutical composition, preferably in form of a capsule, comprisinga blended mixture comprising

-   -   (a) a therapeutically effective amount of        4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile;    -   (b) a filler or a mixture of one or more, e.g. 1, 2, or 3,        fillers;    -   (c) a disintegrant or a mixture of one or more, e.g. 1, 2 or 3        disintegrants;    -   (d) a lubricant or a mixture of one or more, e.g. 1, 2 or 3        lubricants, and    -   (e) a glidant or a mixture of one or more, e.g. 1, 2 or 3        glidants.

In another aspect, the present invention is directed to a method forpreparing a pharmaceutical composition according to the presentinvention comprising the steps of:

-   -   (a) sequentially adding two portions of a filler or a mixture of        fillers and one portion of the therapeutic compound        4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile        (CGP47645) in the order first portion filler, therapeutic        compound, second portion filler into a suitable mixing vessel        and mixing the layers of the components using a suitable mixer        to produce a pre-mix,    -   (b) sieving the pre-mix through a sieve with a mesh size of not        more than 1.0 mm, preferably a mesh size of 0.5 mm,    -   (c) optionally mixing the sieved pre-mix using a suitable mixer,    -   (d) sieving a mixture of the remaining excipients, except the        lubricant, through a sieve with a mesh size of not more than 1.0        mm, preferably a mesh size of 0.5 mm,    -   (e) mixing the pre-mix with the mixture of the remaining        excipients, except the lubricant, using a suitable mixer,    -   (f) sieving the resulting mixture through a sieve with a mesh        size of not more than 1.0 mm, preferably a mesh size of 0.5 mm,    -   (g) optionally mixing the sieved mixture using a suitable mixer,    -   (h) sieving the lubricant, through a sieve with a mesh size of        not more than 1.0 mm, preferably a mesh size of 0.5 mm and        subsequently adding the lubricant to the resulting mixture of        step (f) or (g),    -   (i) mixing the resulting mixture using a suitable mixer to        obtain the final blend,    -   (j) filling the final blend of step (i) into capsules,        optionally using a suitable capsule filling machine.

In a further aspect, the present invention provides the pharmaceuticalcompositions according the present invention for use in the treatment orprevention of a condition or disorder associated with aromataseactivity, including, but not limited to estrogen dependent diseases,such as breast tumor, endometriosis, uterine fibroids, uterineleiomyoma, uterine adenomyosis, dysfunctional uterine bleeding andabnormal endometrial thickening; premature labour; endometrial tumors inwomen; or gynaecomastia in men.

In one embodiment the present invention is directed to thepharmaceutical compositions according to the present invention for usein the treatment of a male patient in need of increased testosteronelevels, preferably an overweight or obese male patient in need ofincreased testosterone levels.

In another embodiment the present invention is directed to thepharmaceutical compositions according to the present invention for usein the treatment of hypogonadism or hypogonadotropic hypogonadism in amale patient, preferably an overweight or obese male patient.

DETAILED DESCRIPTION OF THE INVENTION

There are provided novel pharmaceutical compositions that comprise4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645). The pharmaceutical compositions are in the form of solidoral dosage forms, especially capsules with an immediate releaseprofile. The pharmaceutical compositions may be prepared by preparing ablended mixture comprising4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645) and pharmaceutically acceptable excipients, wherein saidblended mixture subsequently is filled into capsules e.g. by using anencapsulating machinery.

Any capsules as known in the art may be used to encapsulate the blendedmixture. An example of such a capsule are hard gelatin capsules, forexample CONI-SNAP manufactured by Capsugel of Morris Plains, N.J.Suitable sizes for such capsules include, but are not limited to sizesNos. 0 through 5.

Pharmaceutical compositions, in particular in the form of capsules,according to the present invention may contain, for example, from about0.01 mg to up to 20 mg, preferably from about 0.01 mg to about 10 mg,most preferably from about 0.01 to about 5 mg of the therapeuticcompound per capsule; e.g. about 0.01 mg, about 0.05 mg, about 0.1 mg,about 0.3 mg, about 0.5 mg, about 1 mg, about 2 mg, about 3 mg, about 5mg, about 10 mg or about 20 mg therapeutic compound per capsule.

As used herein, the term “therapeutic compound” refers to4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645) of formula I:

As used herein the term “pharmaceutical composition” means, for example,a mixture containing a specified amount of a therapeutic compound, e.g.a therapeutically effective amount, in a pharmaceutically acceptablecarrier to be administered to a mammal, e.g., a human in order to treata disease.

As used herein the term “pharmaceutically acceptable” refers to thosecompounds, materials, compositions and/or dosage forms, which are,within the scope of sound medical judgment, suitable for contact withthe tissues of mammals, especially humans, without excessive toxicity,irritation, allergic response and other problem complicationscommensurate with a reasonable benefit/risk ratio.

As used herein the term “immediate-release” refers to the rapid releaseof the majority of the therapeutic compound, e.g., greater than about50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%,or about 90% within a relatively short time, e.g., within 1 hour, 40minutes, 30 minutes or 20 minutes after oral ingestion. Particularlyuseful conditions for immediate-release are release of at least or equalto about 80% of the therapeutic compound within thirty minutes afteroral ingestion. The particular immediate-release conditions for aspecific therapeutic compound will be recognized or known by one ofordinary skill in the art. The immediate release profile can bedetermined from an in vitro dissolution test.

As used herein the term “excipient” refers to a pharmaceuticallyacceptable ingredient that is commonly used in the pharmaceuticaltechnology for preparing solid oral dosage formulations. Examples ofcategories of excipients include, but are not limited to, binders,disintegrants, lubricants, glidants, stabilizers, fillers and diluents.The amount of each excipient used may vary within ranges conventional inthe art. The following references which are all hereby incorporated byreference disclose techniques and excipients used to formulate oraldosage forms. See The Handbook of Pharmaceutical Excipients, 4thedition, Rowe et al., Eds., American Pharmaceuticals Association (2003);and Remington: the Science and Practice of Pharmacy, 20th edition,Gennaro, Ed., Lippincott Williams & Wilkins (2000).

The term “prevention” refers to prophylactic administration to a healthysubject to prevent the development of a condition. Moreover, the term“prevention” means prophylactic administration to patients being in apre-stage of a condition to be treated.

The term “treatment” is understood the management and care of a patientfor the purpose of combating a disease, condition or disorder.

The present invention provides a low-dose pharmaceutical composition,preferably in form of a capsule, comprising a blended mixture comprising

-   -   (a) a therapeutically effective amount of        4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile;    -   (b) a filler or a mixture of one or more, e.g. 1, 2, or 3        fillers;    -   (c) a disintegrant or a mixture of one or more, e.g. 1, 2 or 3        disintegrants;    -   (d) a lubricant or a mixture of one or more, e.g. 1, 2 or 3        lubricants, and    -   (e) a glidant or a mixture of one or more, e.g. 1, 2 or 3        glidants.

A therapeutically effective amount of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile may befor example from about 0.01 mg to up to 20 mg, preferably from about0.01 mg to about 10 mg, most preferably from about 0.01 to about 5 mgper capsule; e.g. about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.3mg, about 0.5 mg, about 1 mg, about 2 mg, about 3 mg, about 5 mg, about10 mg or about 20 mg per capsule.

In one embodiment, the pharmaceutical composition, in particular in theform of capsules, according to the present invention contains from about0.1 mg to about 10 mg of the therapeutic compound per capsule,preferably from about 0.1 mg to about 5 mg of the therapeutic compoundper capsule.

The fillers to be employed in accordance with the present inventioninclude, without limitation, microcrystalline cellulose (e.g., celluloseMK GR and products available under the registered trade marks AVICEL,FILTRAK, HEWETEN or PHARMACEL, Vivapur, emcocel, tabulose);low-substituted hydroxypropyl cellulose; hydroxyethyl cellulose;hydroxypropyl methyl cellulose; carbohydrates, such as sugars, sugaralcohols, starches or starch derivatives, for example sucrose, lactose,dextrose, saccharose, glucose, sorbitol, mannitol, xylitol, potatostarch, maize starch, rice starch, wheat starch or amylopectin;tricalcium phosphate; calcium hydrogen phosphate; calcium sulfate;dibasic calcium phosphates; magnesium oxide or mixtures thereof.

In some embodiments the filler suitable for use in accordance with thepresent invention has disintegrant properties. Fillers with disintegrantproperties include, without limitation, maize starch, microcrystallinecellulose, alginic acid and pregelatinized starch or mixtures thereof.

Preferably the filler is selected from the group consisting ofmicrocrystalline cellulose, lactose, maize starch and mixtures thereof.

Preferably the lactose is lactose monohydrate. In some embodiments saidlactose monohydrate is spray dried.

More preferably the filler is a mixture of lactose monohydrate,microcrystalline cellulose and maize starch.

The filler may be employed in an amount ranging from about 50% to about99%, preferably from about 70% to about 97%, most preferably from about80-96%, e.g, 85-95% or 89-94% by weight of the capsule content.

In one embodiment, the filler is a combination of lactose monohydrateand two other fillers, e.g. microcrystalline cellulose and maize starch.In one aspect of this embodiment the filler is a mixture of lactosemonohydrate, microcrystalline cellulose and maize starch, wherein theamount of lactose monohydrate in said mixture is between about 60% andabout 75% by weight of the mixture of fillers, preferably between about65% and 70% by weight of the mixture of fillers; the amount ofmicrocrystalline cellulose in said mixture is between about 10% andabout 30% by weight of the mixture of fillers, preferably between about15% and about 25% by weight of the mixture of fillers; the amount ofmaize starch in said mixture is between about 5% and about 20% by weightof the mixture of fillers, preferably between about 7.5% and about 17.5%by weight of the mixture of fillers.

Examples of disintegrants to be employed in accordance with the presentinvention include, without limitation, carboxymethylcellulose calcium(CMC-Ca), carboxymethylcellulose sodium (CMC-Na) or crosscarmellosesodium, e.g. AC-DI-SOL, Sodium Starch Glycolate (SSG), alginic acid,sodium alginate and guar gum or mixtures thereof; preferablycrosscarmellose sodium, e.g. AC-DI-SOL, cross-linked polyvinylpyrrolidone (e.g. CROSPOVIDONE, POLYPLASDONE or KOLLIDON XL) and SodiumStarch Glycolate (SSG) or mixtures thereof.

A preferred disintegrant is Sodium Starch Glycolate (SSG).

The disintegrant may be employed in an amount ranging from about 1% toabout 10%, preferably from about 2% to about 7.5%, most preferably fromabout 3% to about 6%, by weight of the capsule content.

The lubricants to be employed in accordance with the present inventioninclude, without limitation, magnesium stearate, aluminum or calciumsilicate, stearic acid, cutina, PEG 4000-8000, talc or mixtures thereof,preferably sodium stearyl fumarate or magnesium stearate, morepreferably magnesium stearate.

The lubricant may be employed in an amount ranging from about 0.1% toabout 10%, preferably from about 0.25% to about 5%, e.g. about 0.5% toabout 2%, by weight of the capsule content.

Suitable glidants than can be used in accordance with the presentinvention include, without limitation, colloidal silicon dioxide (e.g.,Aerosil 200), magnesium trisilicate, powdered cellulose, starch, talc ormixtures thereof, preferably colloidal silicon dioxide.

The glidant may be employed in an amount ranging from about 0.05% toabout 5%, preferably from about 0.1% to about 1%, more preferably fromabout 0.25% to about 1%, e.g. 0.25%, 0.5% or 0.75%, by weight of thecapsule content.

In one embodiment of the present invention the blended mixture which maybe filled into the capsule comprises

-   -   (a) a therapeutically effective amount of        4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile,    -   (b) a mixture of 3 fillers, wherein said fillers are        microcrystalline cellulose, lactose monohydrate and maize        starch,    -   (c) a disintegrant, wherein said disintegrant is Sodium Starch        Glycollate (SSG),    -   (d) a lubricant, wherein said lubricant is magnesium stearate,    -   (e) a glidant, wherein said glidant is colloidal silicon dioxide        (e.g., Aerosil 200).

In a particular preferred embodiment the blended mixture which may befilled into the capsule comprises

-   -   (a) a therapeutically effective amount of        4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile,    -   (b) a mixture of 3 fillers, wherein said fillers are        microcrystalline cellulose, lactose monohydrate and maize        starch,    -   (c) a disintegrant, wherein said disintegrant is Sodium Starch        Glycollate (SSG),    -   (d) a lubricant, wherein said lubricant is magnesium stearate,    -   (e) a glidant, wherein said glidant is colloidal silicon dioxide        (e.g., Aerosil 200),

-   wherein said therapeutically effective amount of    4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile is    from about 0.01 mg to up to 20 mg, preferably from about 0.01 mg to    about 10 mg, most preferably from about 0.01 mg to about 5 mg per    capsule; e.g. about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.3    mg, about 0.5 mg, about 1 mg, about 2 mg, about 3 mg, about 5 mg,    about 10 mg or about 20 mg per capsule;

-   said mixture of fillers is employed in an amount ranging from about    50% to about 99%, preferably from about 70% to about 97%, most    preferably from about 80% to to about 96%, e.g, 85-95% or 89-94%, by    weight of the capsule content;

-   said disintegrant is employed in an amount ranging from about 1% to    about 10%, preferably from about 2% to about 7.5%, most preferably    from about 3% to about 6%, e.g. 4% or 5%, by weight of the capsule    content;

-   said lubricant is employed in an amount ranging from about 0.1% to    about 10%, preferably from about 0.25% to about 5%, e.g. 0.5%, 1% or    2%, by weight of the capsule content and said glidant is employed in    an amount ranging from about 0.05% to about 5%, preferably from    about 0.1% to about 1%, more preferably from about 0.25% to about    1%., e.g. 0.25%, 0.5% or 0.75%, by weight of the capsule content.

Another aspect of the present invention relates to a method forpreparation a low-dose capsule formulation of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645).

Accordingly there is also provided a method for preparing a low-dosepharmaceutical composition, in the form of a capsule, as describedherein above comprising the steps of:

-   -   (a) sequentially adding two portions of a filler or a mixture of        fillers and one portion of the therapeutic compound        4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile        (CGP47645) in the order first portion filler, therapeutic        compound, second portion filler into a suitable mixing vessel        and mixing the layers of the components using a suitable mixer        to produce a pre-mix;    -   (b) sieving the pre-mix through a sieve with a mesh size of not        more than 1.0 mm, preferably a mesh size of 0.5 mm;    -   (c) optionally mixing the sieved pre-mix using a suitable mixer;    -   (d) sieving a mixture of the remaining excipients, except the        lubricant, through a sieve with a mesh size of not more than 1.0        mm, preferably a mesh size of 0.5 mm;    -   (e) mixing the pre-mix with the mixture of the remaining        excipients, except the lubricant, using a suitable mixer;    -   (f) sieving the resulting mixture through a sieve with a mesh        size of not more than 1.0 mm, preferably a mesh size of 0.5 mm;    -   (g) optionally mixing the sieved mixture using a suitable mixer;    -   (h) sieving the lubricant, through a sieve with a mesh size of        not more than 1.0 mm, preferably a mesh size of 0.5 mm and        subsequently adding the lubricant to the resulting mixture of        step (f) or (g);    -   (i) mixing the resulting mixture using a suitable mixer to        obtain the final blend;    -   (j) filling the final blend of step (i) into capsules,        optionally using a suitable capsule filling machine;

In one embodiment of the present invention the filler used for thepre-mix is maize starch or lactose monohydrate or a mixture thereof,preferably maize starch.

In one embodiment the whole amount of maize starch or lactosemonohydrate is used for the pre-mix. In another embodiment only a partof the maize starch or the lactose monohydrate is used for the premix.

Preferably the filler for the pre-mix is maize starch and the amount ofmaize starch in the pre-mix is between about 5% to about 100% by weightof the total amount of maize starch in the blend, such as for exampleabout 13%, about 70% or about 100% by weight of the total amount ofmaize starch in the blend.

In one embodiment of the present invention the two portions of filler instep (a) are used in a ratio of about 1:1.

In another embodiment steps (c) and/or (g) are mandatory steps.

As mentioned above, formulation of low dose medicines can be verychallenging and problems related to content uniformity and physicalstability may arise. Content uniformity is a key parameter for oralsolid dosage forms, because significant deviations in active content mayimpact the performance of the product in terms of efficacy and safety.The selection of the excipients and the specific steps during themanufacturing are critical factors and need to be controlled in order toget a homogenous and segregation-free low dose formulation.

It has been found that the blend uniformity of the final blended mixtureof the low-dose capsule formulations of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645) of the present invention is between 95% and 101% versus thetheoretical amount. Similarly, the content uniformity after the fillingof the blend into the capsules is between 97% and 104% versus the label.No trend towards segregation was observed (Example 3).

Accordingly, in one embodiment of the invention the blend uniformity ofthe low dose capsule formulation of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645) as described herein above is of between 90% and 110%,preferably of between 95% and 105% versus the theoretical amount.

In another embodiment of the invention the content uniformity of the lowdose capsule formulation of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645) as described herein above is of between 90% and 110%,preferably of between 95% and 105% versus the theoretical amount.

In humans, the compound4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645) formulated in the pharmaceutical composition according to thepresent invention was initially studied in a single, ascending doseprotocol in human female volunteers to assess safety and tolerabilityand pharmacokinetic (PK) and pharmacodynamic (PD) effects of singledoses of CGP47645 (see Example 4).

The study showed that the median T_(max) occurred within 1 hour ofingestion, and that the half life was extremely long, approximately 25days at doses above 0.01 mg.

In one aspect the invention relates to the pharmaceutical compositionsas described herein above for use in the treatment or prevention of acondition or disorder associated with aromatase activity, including, butnot limited to estrogen dependent diseases, such as breast tumor,endometriosis, uterine fibroids, uterine leiomyoma, uterine adenomyosis,dysfunctional uterine bleeding and abnormal endometrial thickening;premature labour; endometrial tumors in women; or gynaecomastia in men.

In one embodiment the invention relates to the pharmaceuticalcompositions as described herein above for use in the treatment of amale patient in need of increased testosterone levels, preferably anoverweight or obese male patient in need of increased testosteronelevels.

In another embodiment the invention relates to the pharmaceuticalcompositions as described herein above for use in the treatment ofhypogonadism or hypogonadotropic hypogonadism in a male patient,preferably an overweight or obese male patient.

In another aspect the invention provides a method for treating orpreventing a condition or disorder associated with aromatase activity,including but not limited to estrogen dependent diseases, such as breasttumor, endometriosis, uterine fibroids, uterine leiomyoma, uterineadenomyosis, dysfunctional uterine bleeding and abnormal endometrialthickening; premature labour; endometrial tumors in women; orgynaecomastia in men, comprising administering to an animal, including ahuman patient, in need of such treatment a therapeutically effectiveamount of the pharmaceutical composition according to the presentinvention.

In one embodiment the invention relates to a method for the treatment ofa male patient in need of increased testosterone levels, preferably anoverweight or obese male patient in need of increased testosteronelevels, comprising administering to said patient a therapeuticallyeffective amount of the pharmaceutical composition as described hereinabove.

In another embodiment the invention relates to a method for thetreatment of hypogonadism or hypogonadotropic hypogonadism in a malepatient, preferably an overweight or obese male patient, comprisingadministering to said patient a therapeutically effective amount of thepharmaceutical composition as described herein above.

The present invention likewise provides the use of a pharmaceuticalcomposition according to the present invention for the manufacturing ofa medicament for the treatment or prevention of a condition or disorderassociated with aromatase activity, including, but not limited toestrogen dependent diseases, such as breast tumor, endometriosis,uterine fibroids, uterine leiomyoma, uterine adenomyosis, dysfunctionaluterine bleeding and abnormal endometrial thickening; premature labour;endometrial tumors in women; or gynaecomastia in men.

In one embodiment the invention relates to the use of a pharmaceuticalcomposition as described herein above for the manufacturing of amedicament for the treatment of a male patient in need of increasedtestosterone levels, preferably an overweight or obese male patient inneed of increased testosterone levels.

In another embodiment the invention relates to the use of apharmaceutical composition as described herein above for themanufacturing of a medicament for the treatment of hypogonadism orhypogonadotropic hypogonadism in a male patient, preferably anoverweight or obese male patient.

The following examples are illustrative, but do not serve to limit thescope of the invention described herein. The examples are meant only tosuggest a method of practicing the present invention.

EXAMPLES Example 1 Preparation of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile

The following example describes a method for the synthesis of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile (alsoknown as4-[α-4-Cyanophenyl)-α-fluoro-1-1,2,4-triazolyl)-methyl]-benzonitrile orCGP47645) as disclosed within Lang et al., U.S. Pat. No. 5,637,605:

A solution of 0.8 mmol of potassium hexamethyldisilazane in 1.6 ml oftoluene is diluted with 5 ml of THF and, after cooling to −78° C., asolution of 190 mg of4-[α-(4-cyanophenyl)-1-(1,2,4-triazolyl)methyl]-benzonitrile (seeEP-A-236 940, Ex. 20a) in 3 ml of THF is added thereto. After stirringfor 1 hour at the same temperature, there are added dropwise to thedark-red solution 301 mg of N-fluoro-dimethylsaccharinsultam in 3 ml ofTHF. After a further 1.5 hours at −78° C., the reaction mixture isheated to room temperature within 1 hour and poured onto a saturatedsolution of ammonium chloride in water and then extracted with methylenechloride. Drying over magnesium chloride and concentration of thesolvent by evaporation yields the crude product which is purified bymeans of flash-chromatography (SiO₂, hexane/ethyl acetate 9:1, 4:1 to1:1). TLC (SiO₂, CHCl₃/methanol 9:1, Rf=0.85); IR (KBr): 2220 cm⁻¹;¹H-NMR (CDCl₃): δ (ppm)=7.46 and 7.76 (8H,m), 8.07 (1H,s), 8.16 (1H,s).

All disclosure relevant to the preparation of4-[α-4-Cyanophenyl)-α-fluoro-1-1,2,4-triazolyl)-methyl]-benzonitriledescribed in Lang et al., U.S. Pat. No. 5,376,669 is hereby incorporatedby reference herein.

The above paragraph refers to EP-A-236 940, Ex. 20a. The U.S. equivalentto EP-236 940 is Bowman, U.S. Pat. No. 4,749,713. Example 20 (a) ofEP-A-236 940 (U.S. Pat. No. 4,749,713) states that4-[1-(1,2,4-Triazolyl)-methyl]-benzonitrile is reacted with potassiumtert-butoxide and 4-fluorobenzonitrile according to the procedure inExample 2 of U.S. Pat. No. 4,749,713 to yield4-[α-(4-cyanophenyl)-1-(1,2,4-triazolyl)-methyl]benzonitrile, m.p. 181°C.-183° C.

The procedure of Example 2 of U.S. Pat. No. 4,749,713 provides that: Asuspension of potassium tert-butoxide (61.6 g) in dimethylformamide (500mL) is stirred and cooled to −10° C. (ice-salt bath), and a solution of4-(1-imidazolylmethyl)-benzonitrile (45.6 g) in dimethylformamide (250mL) is added so that the reaction temperature remains below 0° C. Theresulting solution is stirred at 0° C. for 0.5 hour and then a solutionof 4-fluorobenzonitrile (38.3 g) in dimethylformamide (100 mL) is addedwhile keeping reaction temperature below 5° C. After 0.75 hour, thereaction mixture is neutralized to pH 7 by addition of sufficient 3Nhydrochloric acid and the bulk of the solvents are then removed underreduced pressure. The residue is diluted with water (500 mL) and thecrude product is extracted into ethyl acetate (3×200 mL). The combinedextracts are then extracted with 3N hydrochloric acid (3×150 mL) and,after washing the latter acid extracts with ethyl acetate (100 mL), thesolution is made basic (pH 8) with 6N ammonium hydroxide and the productis again extracted into ethyl acetate (3×150 mL). The combined extractsare dried (MgSO4), decolorized by treatment with charcoal, and thenevaporated to give crude4-[α-(4-cyanophenyl)-1-imidazolylmethyl]-benzonitrile as an oil. Thismaterial is dissolved in isopropanol (250 mL) and the warm solution isstirred with succinic acid (14.4 g). Upon dilution with diethyl ether(100 mL) and stirring at ambient temperature, the hemi-succinate saltseparates. The salt is filtered off, washed with a little coldisopropanol and then air dried to afford4-[α-(4-cyanophenyl)-1-imidazolylmethyl]-benzonitrile hemisuccinate,m.p. 149° C.-150° C. The hemifumarate salt has m.p. 157° C.-158° C.

All disclosure relevant to the preparation of4-[α-(4-cyanophenyl)-1-(1,2,4-triazolyl)-methyl]benzonitrile describedin Bowman, U.S. Pat. No. 4,749,713 is hereby incorporated by referenceherein.

Example 2 Low-Dose Capsule Formulations of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645)

CGP47645 containing hard gelatine capsules are prepared by the followingprocess: The required excipients, in the respective amounts to yield thefinal composition as indicated in Table 1 below, and the appropriateamount of CGP47645 drug substance are weighed.

Then, approximately 50% of corn starch is filled into suitablecontainer, the drug substance is added, followed by the remaining 50% ofcorn starch to get a sandwich of drug substance between two layers ofmaize starch. Blending and sieving this mixture yields the drugsubstance (DS) premix.

The remaining excipients (microcrystalline cellulose, spray-driedlactose, sodium starch glycolate, and colloidal silicon dioxide[Aerosil® 200]) are mixed and sieved and transfer into a suitablecontainer. Then the DS premix is added into container containing thesieved excipients and the mixture is blended together. Finally,pre-sieved Magnesium stearate is added to the blend containing the DSand this mixture is blended again to yield the final blend. The finalblend is filled into hard gelatin capsules.

All the excipients comply with the requirements of the applicablecompendial monographs (Ph.Eur., NF). The hard gelatine capsules arepackaged in HDPE bottles with aluminum induction seal equipped withchild-resistant screw-cap closures.

The final dosage form is a hard gelatine capsule containing a white toyellowish powder in a pink opaque capsule, size 1 or 3.

The following Table 1 indicates the composition of the CGP47645 hardgelatin capsule of 0.1 mg, 0.5, 1 mg and 10 mg strength.

TABLE 1 Amount per capsule (mg) Ingredient 0.1 mg¹ 0.1 mg² 0.5 mg¹ 1 mg²10 mg² Capsule content CGP47645 0.1 0.1 0.5 1.0 10.0 Lactose monohydrate96.0 192.0 96.0 192.0 175.5 Cellulose, 30.0 60.0 30.0 60.0 50.0microcrystalline Corn Starch 14.15 28.4 13.75 27.5 40.0 Sodium starchglycolate 7.5 15.0 7.5 15.0 15.0 (Type A) Magnesium Stearate 1.5 3.0 1.53.0 3.0 Silica, colloidal 0.75 1.5 0.75 1.5 1.5 anhydrous Capsule fillweight 150.0 300.0 150 300.0 295.0 Empty capsule shell Capsule shell48.0 76.0 48.0 76.0 76.0 Total capsule weight 198.0 376.0 198.0 376.0371.0 ¹Filled in size 3 capsules; ²Filled in size 1 capsules

Example 3 Formulations of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645)

The required excipients, in the respective amounts to yield the finalcomposition as indicated in Table 2 below, and the appropriate amount ofCGP47645 drug substance are weighed.

Then, approximately 50% of the amount of the filler of the premix, asindicated in Table 2 below, is filled into suitable container, the drugsubstance is added, followed by the remaining 50% of the amount offiller of the premix, as indicated in Table 2, to get a sandwich of drugsubstance between two layers of filler. Blending, sieving and againblending this mixture yields the drug substance (DS) premix.

The remaining excipients, except magnesium stearate, are mixed andsieved and transfer into a suitable container. Then the DS premix isadded into container containing the sieved excipients and the mixture isblended together, sieved and mixed again. Finally, pre-sieved Magnesiumstearate is added to the blend containing the DS and this mixture isblended again to yield the final blend. The final blend is filled intohard gelatin capsules.

All the excipients comply with the requirements of the applicablecompendial monographs (Ph.Eur., NF). The hard gelatine capsules arepackaged in HDPE bottles with aluminum induction seal equipped withchild-resistant screw-cap closures.

The final dosage form is a hard gelatine capsule containing a white toyellowish powder in a pink opaque capsule, size 1 or size 3.

The following Table 2 indicates the composition of the CGP47645 hardgelatin capsule of 0.1 mg and 0.5 mg strength.

TABLE 2 Amount per capsule (mg) (Trial No.) 0.5 0.5 0.5 0.1 0.1 0.5Ingredient mg mg mg mg mg mg Capsule content (1) (2) (3) (4) (5) (6) DSpremix CGP47645 0.5 0.5 0.5 0.1 0.1 0.5 Corn Starch 9.5 13.75 — 14.1514.15 13.75 Lactose — — 13.75 — — — monohydrate Corn Starch 4.25 — 13.75— — — Cellulose, 30.0 30.0 30.0 30.0 30.0 30.0 microcrystalline Lactose96.0 96.0 82.25 96.0 96.0 96.0 monohydrate Sodium starch 7.5 7.5 7.5 7.57.5 7.5 glycolate (Type A) Magnesium 1.5 1.5 1.5 1.5 1.5 1.5 StearateSilica, colloidal 0.75 0.75 0.75 0.75 0.75 0.75 anhydrous Capsule fillweight 150.0 150.0 150.0 150.0 150.0 150.0

Blend Uniformity and Content Uniformity of the blended mixtures orcapsules according to Table 2 are determined by HPLC analysis of tensamples of each composition.

The following Table 3 shows the average values of blend uniformity (BU)and content uniformity (CU) at the end of the filling step of theblended mixtures of Table 2 or the corresponding hard gelatin capsules,respectively.

TABLE 3 Amount per capsule (mg) (Trial No.) 0.5 0.5 0.5 0.1 0.1 0.5 mgmg mg mg mg mg (1) (2) (3) (4) (5) (6) BU 98.7 95.5 98.7 100.1 96.6 99.4(vs. theoretical amount) CU at the end of 99.8 99.2 101.2 103.5 97.0102.3 the filling step (% vs. label)

Blend uniformity of the final blends according to Example 3 are between95% and 101% versus the theoretical amount. Similarly the contentuniformity after filling of the blend into the capsules is between 97%and 104% versus the label. No trend towards segregation was observed.

Example 4 Single ascending dose study of4,4′-[Fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645)

This was a randomized, double-blind, placebo- and active-controlledsingle ascending dose study in pre- and post-menopausal women to assessthe safety and tolerability, PK and PD effects of single doses of4,4′-[Fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645). There were 8 cohorts of 8 post-menopausal subjectsrandomized 6:2, CGP47645:placebo, who received single doses of CGP47645beginning at the dose of 0.01 mg and carried through 20 mg, whichreached the limit of the toxicology exposure coverage. Patients receivedeither 0.1 mg, 1 mg, and 10 mg drug substance containing hard gelatincapsules as described in Example 2 or appropriate matching placebocapsules. For the lowest two dosing cohorts, 0.1 mg drug containingcapsules were used for reconstituting the CGP47645 oral solutions fordosing the 0.01 and 0.03 dosing strength (Cohort 1 and 2).

A minimal toxic dose (MTD) was not reached. A single cohort of 8pre-menopausal subjects without childbearing potential (Cohort No. 9)received CGP47645 0.1 mg or placebo, randomized 6:2, and one last cohortreceived letrozole 2.5 mg as an internal positive control cohort for thePD measurements. Table 4 presents the PK parameters based on preliminaryanalysis of the concentration-time profile obtained from this study.

TABLE 4 CGP47645 Pharmacokinetics in Post- & Pre-menopausal womenC_(max) AUC(0-t_(last)) Dose Cohort (ng/mL) T_(max) (hr) (ng*hr/mL)T_(1/2) (days) (mg) No. Size Mean CV (%) Median Mean CV (%) Mean CV (%)0.01 1 (n = 5) 0.2 21.7 1 1.4 53.2 2.3 127.5 0.03 2 (n = 6) 0.4 18.7 0.624.1 34.0 16.5 36.0 0.1 3 (n = 6) 1.8 13.4 1 123.1 10.7 18.2 10.9 0.3 4(n = 6) 5.1 14.1 1 605.1 49.0 23.5 19.9 1 5 (n = 5) 12.8 22.0 1 3201.937.2 22.4 38.5 3 6 (n = 6) 38.4 17.0 1 10053.0 16.7 25.0 8.4 10 7 (n =6) 123.8 26.4 2 41745.5 17.3 27.3 17.6 20 8 (n = 6) 269.8 30.9 2 76731.611.4 26.9 16.5 0.1 9 (n = 6) 1.7 15.1 1 116.2 17.1 23.5 31.0 2.5Letrozole (n = 8) 33.5 27.0 1 1667.7 40.8 2.9 40.7

CGP47645 exhibited dose proportional pharmacokinetics and adose-dependent inhibition of estrone, estrone sulfate and estradiol. Nodifferences in CGP47645 pharmacokinetics were observed between post- andpre-menopausal women. CGP47645 is rapidly absorbed with a T_(max) of0.5-2 hrs. Both C_(max) & AUC increased in a dose-proportional manner.CGP47645 exhibited low inter-subject variability of 10-30% andcompletely unexpected long half-life in the range of 23 to 27 days.

The study showed evidence of efficacy in PD parameters with estronesuppression at least equal to letrozole already at doses of 0.1 mg and0.3 mg. In postmenopausal women, the lowest single dose at whichtransient estrogen suppression was seen was 0.01 mg; and the lowestsingle dose at which maximal estrogen suppression was observed inpost-menopausal women, using chemiluminescence or radioimmunoassay, was0.1 mg. No inhibition of other enzymes involved in steroid hormonesynthesis or metabolism was observed; in particular there were nochanges in androgen levels, progesterone, aldosterone, cortisol, ACTH,or 17-keto or 17-OH steroids in 24 hour urine collections. There were nochanges in bone density by DEXA after 6 months.

1. A low-dose pharmaceutical composition, comprising a blended mixturecomprising (a) a therapeutically effective amount of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile; (b) oneor more filler; (c) one or more disintegrant; (d) one or more lubricant,and (e) one or more glidant.
 2. A pharmaceutical composition accordingto claim 1, wherein said therapeutically effective amount of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile is fromabout 0.01 mg to up to 20 mg per dosage unit capsule.
 3. Apharmaceutical composition according to claim 1, wherein said filler isselected from one or more of microcrystalline cellulose; low-substitutedhydroxypropyl cellulose; hydroxyethyl cellulose; hydroxypropyl methylcellulose; carbohydrates, such as sugars, sugar alcohols, starches ofstarch derivatives, for example sucrose, lactose, dextrose, saccharose,glucose, sorbitol, mannitol, xylitol, potato starch, maize starch, ricestarch, wheat starch or amylopectin; tricalcium phosphate, calciumhydrogen phosphate, calcium sulfate, dibasic calcium phosphates, andmagnesium oxide.
 4. A pharmaceutical composition according to claim 1,wherein said filler is selected from one or more of microcrystallinecellulose, lactose, and maize starch.
 5. A pharmaceutical compositionaccording to claim 1, wherein said disintegrant is selected from one ormore of carboxymethylcellulose calcium (CMC-Ca), carboxymethylcellulosesodium (CMC-Na), crosscarmellose sodium, Sodium Starch Glycolate,alginic acid, sodium alginate and guar gum.
 6. A pharmaceuticalcomposition according to claim 1, wherein said lubricant is selectedfrom one or more of magnesium stearate, aluminium silicate, calciumsilicate, stearic acid, cutina, PEG 4000-8000, and talc.
 7. Apharmaceutical composition according to claim 1, wherein said glidant isselected from one or more of colloidal silicon dioxide, magnesiumtrisilicate, powdered cellulose, starch, and talc.
 8. A method forpreparing a pharmaceutical composition according to claim 1, comprisingthe steps of: (a) sequentially adding two portions of one or more fillerand one portion of the therapeutic compound4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile(CGP47645) in the order first portion filler, therapeutic compound,second portion filler into a suitable mixing vessel and mixing thelayers of the components using a suitable mixer to produce a pre-mix,(b) sieving the pre-mix through a sieve with a mesh size of not morethan 1.0 mm, preferably a mesh size of 0.5 mm, (c) optionally mixing thesieved pre-mix using a suitable mixer, (d) sieving a mixture of theremaining excipients, except the lubricant, through a sieve with a meshsize of not more than 1.0 mm, preferably a mesh size of 0.5 mm, (e)mixing the pre-mix with the mixture of the remaining excipients, exceptthe lubricant, using a suitable mixer, (f) sieving the resulting mixturethrough a sieve with a mesh size of not more than 1.0 mm, (g) optionallymixing the sieved mixture using a suitable mixer, (h) sieving thelubricant, through a sieve with a mesh size of not more than 1.0 mm, andsubsequently adding the lubricant to the resulting mixture of step (f)or (g), (i) mixing the resulting mixture using a suitable mixer toobtain the final blend, and (j) filling the final blend of step (i) intocapsules, optionally using a suitable capsule filling machine.
 9. Amethod of treating a condition or disorder associated with aromataseactivity, comprising administering a therapeutically effective amount toa subject in need thereof a pharmaceutical composition according toclaim
 1. 10. The method according to claim 9, wherein said condition ordisorder associated with aromatase activity is selected from one or moreof i. one or more estrogen dependent disease, breast tumor,endometriosis, uterine fibroids, uterine leiomyoma, uterine adenomyosis,dysfunctional uterine bleeding and abnormal endometrial thickening;premature labour; endometrial tumors in women; gynaecomastia in men; ii.low testosterone in men; and iii. hypogonadism or hypogonadotropichypogondadism in a male patient.
 11. The method of claim 10 wherein saidtherapeutically effective amount of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile is fromabout 0.01 mg to about 20 mg per dosage unit capsule.
 12. Thepharmaceutical composition of claim 2 wherein said therapeuticallyeffective amount of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile is fromabout 0.01 mg to about 10 mg per dosage unit.
 13. The pharmaceuticalcomposition of claim 12 wherein said therapeutically effective amount of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile is fromabout 0.01 mg to about 5 mg per dosage unit.
 14. The pharmaceuticalcomposition of claim 5 wherein said disintegrant is Sodium StarchGlycolate.
 15. The pharmaceutical composition of claim 6 wherein saidlubricant is magnesium stearate.
 16. The pharmaceutical composition ofclaim 6 wherein said glidant is colloidal silicon dioxide.
 17. Themethod of claim 10 wherein said condition is hypogonadism orhypogonadotropic hypogondadism in a male patient.
 18. The method ofclaim 17, wherein said pharmaceutical composition is a capsulecomprising about 0.01 mg to about 3.0 mg of4,4′-[fluoro-(1-H-1,2,4-triazol-1-yl)methylene]bisbenzonitrile.